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Showing content from https://n2t.net/addgene:62987 below:

Addgene: pSpCas9n(BB)-2A-Puro (PX462) V2.0

• Purpose

  Cas9n (D10A nickase mutant) from S. pyogenes with 2A-Puro, and cloning backbone for sgRNA (V2.0)

This material is available to academics and nonprofits only.

• Vector backbone

  PX462
• Total vector size (bp) 9200

• Vector type

  Mammalian Expression, CRISPR

• Selectable markers

  Puromycin

• Bacterial Resistance(s)

  Ampicillin, 100 μg/mL

• Growth Temperature

  37°C

• Growth Strain(s)

  Stbl3

• Copy number

  Unknown

• Gene/Insert name

  hSpCas9n-2A-Puro

• Alt name

  SpCas9n-2A-Puro V2.0

• Alt name

  Cas9n

• Alt name

  PX462 V2.0

• Species

  Synthetic

• Insert Size (bp)

  6000

• Mutation

  Cas9 D10A nickase mutant
• Tags / Fusion Proteins
  • 3XFLAG (N terminal on insert)
  • 2A-Puro (C terminal on insert)

• Supplemental Documents

  • Zhang lab general target sequence cloning protocol

• Addgene Notes

  • Addgene diagnostic digest

• Articles Citing this Plasmid

  • 88 References

• Academic/Nonprofit Terms

  • UBMTA

• Industry Terms

  • Not Available to Industry

• Zeocin® is an InvivoGen trademark.

For more information on Zhang Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/zhang/

The guide sequence is cloned into this plasmid using BbsI sites. Please see the cloning protocol provided by the Zhang lab for more details.

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

• For your Materials & Methods section:

  pSpCas9n(BB)-2A-Puro (PX462) V2.0 was a gift from Feng Zhang (Addgene plasmid # 62987 ; http://n2t.net/addgene:62987 ; RRID:Addgene_62987)

• For your References section:

  Genome engineering using the CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. 10.1038/nprot.2013.143 PubMed 24157548

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